title: Epitope selection to male specific antigens for sex selection in swine.
abstract: Immunological approaches to gender selection have been contemplated since the discovery of the family of male-specific H-Y antigens found only on the surface of male cells. H-Y antigens are able to elicit an immune reaction when cells or tissues from a male donor are grafted to a female recipient. We describe here the development and testing of an inexpensive approach using polyclonal antibodies against four specific H-Y outer membrane proteins male enhanced antigen 1 (MEA 1), male enhanced antigen 2 (MEA 2), sex determining region Y (SRY) and testis determining factor (TDF). Epitopes based on hydrophilic primary sequences of the proteins were synthesized, N-terminal biotin-labeled, linked to streptavidin and mixed with a Ribi adjuvant prior to immunization in rabbits. The antiserum was tested to determine affinity to swine spermatozoa using anti-motility, flow cytometry and motility and sedimentation chambers. Fluorescent microscopy and fluorescent in situ hybridization (FISH) was used to identify the percentage of motile spermatozoa that contained the Y chromosome. We found that the polyclonal antibodies had high affinity to the spermatozoa leading to a cessation of motility. Furthermore, the majority of these non-motile spermatozoa contained the Y chromosome. We conclude that the use of polyclonal antiserum against synthetic H-Y peptide antigens may be an inexpensive and simple means to inhibit the motility of swine spermatozoa bearing the Y chromosome.
authors.affiliation: Department of Biochemistry Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada.
authors.forename: Azarm Akhavien
authors.initials: AA
authors.lastname: Mohammadi
authors.forename: Jason A
authors.initials: JA
authors.lastname: Tetro
authors.forename: Lionel G
authors.initials: LG
authors.lastname: Filion
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology*; D000915:Antibody Affinity; D002465:Cell Movement / Q000187:drug effects; D017358:Cell Wall Skeleton / Q000008:administration & dosage; D002478:Cells, Cultured; D003311:Cord Factors / Q000008:administration & dosage; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000378:metabolism*; D017404:In Situ Hybridization, Fluorescence; D008050:Lipid A / Q000008:administration & dosage / Q000031:analogs & derivatives; D008297:Male; D018761:Multiple Endocrine Neoplasia Type 1 / Q000276:immunology / Q000378:metabolism; D018813:Multiple Endocrine Neoplasia Type 2a / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000378:metabolism; D012743:Sex Preselection / Q000379:methods; D051876:Sex-Determining Region Y Protein / Q000276:immunology / Q000378:metabolism; D013094:Spermatozoa / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D013552:Swine
chemical_list: D000906:Antibodies; D017358:Cell Wall Skeleton; D003311:Cord Factors; D000939:Epitopes; D008050:Lipid A; D010446:Peptide Fragments; C075897:Ribi adjuvant; D051876:Sex-Determining Region Y Protein